System and method for converting adipose derived mesenchymal stems cells to hematopoietic stem/progenitor cells and differentiating into blood cells and applications of same

Abstract

A method of producing hematopoietic stem/progenitor cells (HS/PCs) and blood cells from adipose derived mesenchymal stem cells (Ad MSCs). The method comprises obtaining biological cells of a subject; expanding the biological cells; obtaining a pure Ad MSCs cell line from the expanded biological cells; converting the pure Ad MSCs cell line into HS/PCs in vitro; expanding the converted HS/PCs in vitro; and obtaining the expanded HS/PCs.

What is claimed is: 
     
         1 . A method of producing hematopoietic stem/progenitor cells (HS/PCs) and blood cells from adipose derived mesenchymal stem cells (Ad MSCs), the method comprising:
 obtaining biological cells of a subject;   expanding the biological cells;   obtaining a pure Ad MSCs cell line from the expanded biological cells;   converting the pure Ad MSCs cell line into HS/PCs in vitro;   expanding the converted HS/PCs in vitro; and   obtaining the expanded HS/PCs.   
     
     
         2 . The method according to  claim 1 , the methods further comprising:
 differentiating the expanded HS/PCs into a blood cell line;   wherein the blood cell line comprises at least one of white blood cells (WBC), red blood cells (RBC), and platelets.   
     
     
         3 . The method according to  claim 1 , wherein the step of converting the pure Ad MSCs cell line into the HS/PCs does not use any exogenous genes. 
     
     
         4 . The method according to  claim 1 , wherein the step of obtaining the pure Ad MSCs cell line comprises:
 isolating the pure Ad MSCs cell line from the biological sample; and   culturing the pure Ad MSCs cell line.   
     
     
         5 . The method according to  claim 1 , wherein the step of converting the pure Ad MSCs cell line into the HS/PCs comprises:
 using a HS/PCs converting kit.   
     
     
         6 . The method according to  claim 5 , wherein the HS/PCs converting kit comprises:
 a basic Ad MSCs culture medium;   a basic nutrition supplement; and   a basic converting supplement mixture.   
     
     
         7 . The method according to  claim 6 , wherein the step of converting the pure Ad MSCs cell line into the HS/PCs comprises:
 preparing a HS/PC-C completely medium by mixing the basic nutrition supplement and the basic converting supplement mixture into the basic Ad MSCs culture medium;   adding the HS/PC-C completely medium to the pure Ad MSCs cell line obtained; and   incubating the pure Ad MSCs cell line in the HS/PC-C completely medium for a period of time.   
     
     
         8 . The method according to  claim 5 , wherein the step of converting the pure Ad MSCs cell line into the HS/PCs has a conversion efficiency of more than 10%. 
     
     
         9 . The method according to  claim 5 , wherein the converted HS/PCs have a gene transfection efficiency 25-fold higher than that of regular converted HS/PCs which are not converted by the HS/PCs converting kit. 
     
     
         10 . The method according to  claim 5 , wherein the converted HS/PCs comprises gene carry-on cells. 
     
     
         11 . A HS/PCs converting kit for converting adipose derived mesenchymal stem cells (Ad MSCs) into hematopoietic stem/progenitor cells (HS/PCs), the kit comprising:
 a basic Ad MSCs culture medium;   a basic nutrition supplement; and   a basic converting supplement mixture,   wherein the kit converts the Ad MSCs into HS/PCs.   
     
     
         12 . The kit according to  claim 11 , wherein the basic nutrition supplement comprises fetal bovine serum and horse serum. 
     
     
         13 . The kit according to  claim 11 , wherein the basic converting supplement mixture comprises at least one of insulin, holo-transferrin, sodium selenite (ITS liquid), L-ascorbic acid, GM-CSF, SCF, VEGF, IGF-I, IGF-II, IL-3, Flt3-L, thrombopoietin (TPO), dexamethasone, fatty acid free BSA, 1-thioglycerol, SB431542, CHIR99021, and Y-27632. 
     
     
         14 . The kit according to claim  14 , wherein the basic converting supplement mixture comprises insulin, holo-transferrin, sodium selenite (ITS liquid), L-ascorbic acid, GM-CSF, SCF, VEGF, IGF-I, IGF-II, IL-3, Flt3-L, thrombopoietin (TPO), dexamethasone, fatty acid free BSA, 1-thioglycerol, SB431542, CHIR99021, and Y-27632. 
     
     
         15 . A red blood cells (RBCs) and platelets differentiation kit for differentiating hematopoietic stem/progenitor cells (HS/PCs) into RBCs and platelets, the kit comprising:
 a basic cell culture medium;   a basic nutrition supplement; and   a RBCs supplement mixture;   wherein the kit differentiates the HS/PCs into at least one of RBCs and platelets.   
     
     
         16 . The kit according to  claim 15 , wherein the basic nutrition supplement comprises fetal bovine serum and horse serum. 
     
     
         17 . The kit according to  claim 15 , wherein the RBCs supplement mixture comprises at least one of insulin, holo-transferrin, sodium selenite (ITS liquid), L-ascorbic acid, SCF, VEGF, IGF-I, IGF-II, IL-3, Flt3-L, thrombopoietin (TPO), dexamethasone, fatty acid free BSA, 1-thioglycerol, EPO, ferrous sulfate, PDGF BB, activin A. 
     
     
         18 . A white blood cells (WBCs) differentiation kit for differentiating hematopoietic stem/progenitor cells (HS/PCs) into WBCs, the kit comprising:
 a basic cell culture medium;   a basic nutrition supplement; and   a WBCs supplement mixture;   wherein the kit differentiates the HS/PCs into WBCs.   
     
     
         19 . The kit according to  claim 18 , wherein the basic nutrition supplement comprises fetal bovine serum and horse serum. 
     
     
         20 . The kit according to  claim 18 , wherein the basic converting supplement mixture comprises at least one of insulin, holo-transferrin, sodium selenite (ITS liquid), L-ascorbic acid, SCF, VEGF, IGF-I, IGF-II, IL-3, Flt3-L, thrombopoietin (TPO), dexamethasone, fatty acid free BSA, 1-thioglycerol, BMP4, IL-7, IL-12, IL-11, IL-6, IL-2, FGF-b, TGF-a, TGF-b1, TGF-b2, TGF-b3, IL-1a, IL-1b, IL-4, IL-5, IL-8, IL-10, IL-12, IL-32a, mIL-36R2, GM-CSF. 
     
     
         21 . An automatic multifunctional single cell processor and culture device, the device comprising:
 a housing;   at least one small fluid volume dispenser system disposed inside the housing; and   a tissue grinding system disposed below the small fluid volume dispenser system and in the housing;   wherein the small fluid volume dispenser system is configured to dispense at least one fluid; and wherein the tissue grinding system receives the at least one fluid.   
     
     
         22 . The device according to  claim 21 , wherein the small fluid volume dispenser system is configured to dispense the at least one fluid in a volume between about 5-10 ml. 
     
     
         23 . The device according to  claim 21 , wherein the small fluid volume dispenser system comprises a cylinder houses one or more pre-filled cartridges. 
     
     
         24 . The device according to  claim 23 , wherein the small fluid volume dispenser system comprises a top needle disposed above the one or more pre-filled cartridges and a bottom needle disposed below the one or more pre-filled cartridges. 
     
     
         25 . The device according to  claim 24 , wherein the top needle and the bottom needle are configured to move in a vertical direction for piercing the pre-filled cartridges. 
     
     
         26 . The device according to  claim 21 , wherein the tissue grinding system comprises a vial mechanically connected to a motor. 
     
     
         27 . The device according to  claim 26 , wherein the vial houses a top grinder plate, a bottom grinder plate, a filter disposed below the top grinder plate and the bottom grinder plate. 
     
     
         28 . The device according to  claim 26 , wherein the top grinder plate and the bottom grinder plate are arranged to form a sample disposing place for receiving a biological sample. 
     
     
         29 . The device according to  claim 28 , wherein the tissue grinding system further comprises a tubing fluidly connected to a bottom of the vial on a first end. 
     
     
         30 . The device according to  claim 29 , wherein the tubing fluidly connected to a dispenser funnel on a second end. 
     
     
         31 . The device according to  claim 21 , wherein the device comprises a cellular flask rotator, wherein the cellular flask rotator comprises multiple magnetic nanoparticles.